Mildew Of Cucumber Research Articles

IGDM: Image-Based Grading System of Downy Mildew in Cucumber Using Digital Image Processing and Unsupervised Learning

IGDM: Image-Based Grading System of Downy Mildew in Cucumber Using Digital Image Processing and Unsupervised Learning

Multiplex gene editing reveals cucumber MILDEW RESISTANCE LOCUS O family roles in powdery mildew resistance.

Powdery mildew (PM) is one of the most widespread and prevalent diseases that affects a wide range of crops. In cucumber (Cucumis sativus L.), previous forward genetic studies have identified MILDEW RESISTANCE LOCUS O 8 (CsMLO8) as necessary but alone insufficient for cucumber PM resistance (PMR) and suggested the involvement of other members of the CsMLO family. However, the function of other CsMLO family members in cucumber remains largely unknown. Here, we developed a highly efficient multiplex gene editing system in cucumber to generate a series of Csmlo mutants from all the 13 family members. Systematic analysis of these mutants revealed growth effects of these CsMLO family members on development and PMR. Importantly, we obtained the Csmlo1/8/11 triple mutant with complete resistance to PM. Transcriptome and proteome analysis of PM-resistant Csmlo mutants suggested that the kinesin-like calmodulin-binding protein (KCBP)-interacting Ca2+-binding protein (CsKIC), calmodulin-like protein 28 (CsCML28), and Ca2+-dependent protein kinase 11 (CsCPK11)-mediated calcium signaling pathway is involved in PMR. CsMLO8 interacted directly with CsKIC, and the simultaneous silencing of both genes resulted in a phenotype that resembled the silencing of CsKIC alone. Silencing CsCML28 and CsCPK11 increased susceptibility to PM, whereas overexpressing CsCPK11 through genetic transformation enhanced cucumber's PMR, demonstrating their positive regulatory roles in PMR. Given the importance of PMR for cucurbit crops, this research provides unprecedented insights into the function of the proteins encoded by the CsMLO gene family as well as the plant defense response to PM pathogen.

Molecular genetic basis of resistance to downy mildew in cucumber and melon

Molecular genetic basis of resistance to downy mildew in cucumber and melon

New Insights into the Anti-Oomycete Mechanisms of a Leaf Extract from Glycyrrhiza glabra Against Pseudoperonospora cubensis.

Cucurbit downy mildew, caused by Pseudoperonospora cubensis, is responsible for high economic losses worldwide in cucumber production. Synthetic pesticides or copper-based products are still important tools to manage the disease. However, the pathogen has developed resistance against common fungicides rather quickly, and there is a need for alternative plant-protecting agents. Glycyrrhiza glabra leaf extract is known for its antifungal activity and was highly effective in former bioassays and semi-commercial trials against downy mildew of cucumber. To elucidate the active ingredients and the mode of action, licorice leaf extract was here fractionated into five fractions (F1 to F5) with a newly developed and optimized separation process via flash chromatography. The crude extract (P1) and fraction F1 inhibited the zoospore release from sporangia, the zoospore germination, and germ tube development of P. cubensis almost completely on two cucumber cultivars, one tolerant and one susceptible to the pathogen. Infestation rates were reduced between 73 and 96%. F1 contained three previously reported antifungal polyphenols: glabranin, pinocembrin, and licoflavanon. Here, we report an additional new compound, naringenin. Furthermore, F5 is found here to show some protective effects against P. cubensis, based on either direct fungicidal or indirect effects via the host plant. The presented results show that licorice leaf extract can serve as an alternative plant protection agent, able to manage P. cubensis infestation on cucumber cultivars with differing levels of susceptibility by interfering with important early stages in the pathogen development.

Efficacy of fungicides against powdery mildew of cucumber (Cucumis sativus L.)

Efficacy of fungicides against powdery mildew of cucumber (Cucumis sativus L.)

Defense Mechanisms Induced by Celery Seed Essential Oil against Powdery Mildew Incited by Podosphaera fusca in Cucumber.

This study aimed to evaluate the effectiveness of essential oil extracted from celery (Apium graveolens) seeds (CSEO) for the control of powdery mildew of cucumber (Cucumis sativus) incited by Podosphaera fusca and to investigate the metabolic and genetic defense mechanisms triggered by the treatment with this essential oil in cucumber seedlings. The main compounds in the CSEO as determined by gas chromatography-mass spectrometry (GC-MS) analysis were d-limonene, 3-butyl phthalide, β-selinene, and mandelic acid. The treatment with CSEO led to an increase in the content of both chlorophyll and phenolic/flavonoid compounds in cucumber leaves. In greenhouse tests, the application of CSEO reduced by 60% the disease severity on leaves of cucumber plants and stimulated the activity of defense-related enzymes such as β-1,3-glucanase, chitinase, phenylalanine ammonia-lyase, peroxidase, and polyphenol oxidase. Moreover, treatment with CSEO induced overexpression of β-1,3-glucanase, chitinase, and phenylalanine ammonia-lyase genes. A highly significant correlation was found between the β-1,3-glucanase, chitinase, and phenylalanine ammonia-lyase enzymatic activities and the relative expression of the corresponding encoding genes in both inoculated and non-inoculated cucumber seedlings treated with the essential oil. Overall, this study showed that CSEO is a promising eco-friendly candidate fungicide that can be exploited to control cucumber powdery mildew.

β-Aminobutyric Acid and Powdery Mildew Infection Enhanced the Activation of Defense-Related Genes and Salicylic Acid in Cucumber (Cucumis sativus L.).

Powdery mildew disease, caused by Sphaerotheca fusca, is a major disease affecting cucumbers cultivated in greenhouses. This study was conducted to find defense genes induced by β-aminobutyric acid (BABA) and powdery mildew in cucumber. Disease severities of 25% and 5% were exhibited by the 2000 and 5000 mg/L BABA-treated cucumber, respectively. BABA did not affect the spore germination of the powdery mildew pathogen, showing that BABA is not an antifungal agent against the pathogen. In quantitative real-time PCR analysis, BABA-treated cucumber upregulated the transcriptional levels of the defense genes CsPAL, CsPR3, CsPR1, CsLOX1, CsLOX23, Cs LecRK6.1, CsWRKY20, and Cupi4 in cucumber to maximum levels at 48 h, whereas CsLecRK6.1 reached maximum expression after 24 h, and further, salicylic acid (SA) levels were significantly increased in BABA-treated cucumber plants. In addition, the cucumber infected with powdery mildew underwent a 1.6- to 47.3-fold enhancement in the defense genes PAL, PR3, PR1, Lox1, Lox 23, LecRK6.1, WRKY20, and Cupi4 compared to heathy cucumber. These results suggest that the BABA-induced defense response is associated with SA signaling pathway-dependent systemic acquired resistance (SAR) in cucumber, which is involved in plant resistance mechanisms.

Relative biochemical and physiological docking of cucumber varieties for supporting innate immunity against Podosphaera xanthii

Powdery mildew in cucumber is caused by the Podosphaera xanthii. No strategy for improving disease resistance can be successful in the absence of thorough insights into the physiological and biochemical responses of cucumber plants to powdery mildew. Therefore, a field experiment was executed to evaluate five commercial cucumber varieties (V1: Dynasty, V2: Long green, V3:Desi Kheera, V4:Thamin II, V5:Cucumber 363) for their inherent immunity to powdery mildew. Upon inoculating cucumber plants with Podosphaera xanthii, we noted differential responses among the varieties. Compared to other varieties, V1 and V2 showed higher values (P ≤ 0.05) for chlorophyll-a under control and pathogen-attacked plants respectively. The minimum value of anthocyanin content (−53.73%) was recorded in V3 as compared to other varieties post pathogen infection. All pathogen-infected cucumber varieties showed a considerable (P ≤ 0.05) loss in flavonoid content except V2. The maximum destruction for Phenolics under powdery mildew (179%) were recorded in V4, whereas V1 exhibited maximum phenolic content under control conditions. In pathogen-infected plants, the minimum AsA was recorded in V5 as compared to all other varieties. Pathogen invasion impacted significantly (P ≤ 0.05) the activity of superoxide dismutase (SOD). Besides, cucumber plants after pathogen inoculation resulted in a considerable (P ≤ 0.05) increase of peroxidase (POD) activity in V1 (5.02%), V2 (7.5%), and V3 (11%) in contrast to V4. Our results confirmed that cucumber varieties perform differently, which was brought on by distinct metabolic and physiological modifications that have an impact on growth and development. The changes in different attributes were correlated with cucumber resistance against powdery mildew. The results would help us fully harness the potential of these varieties to trigger disease management initiatives and defense responses.

Diagnosis the Pseudoperonospora cubensis Causes Downy Mildew in Cucumbers and its Resistance to Potassium Silicate and Extract Eggplant Wild

A field experiment was carried out to test the efficiency of potassium silicate and wild eggplant Solanum eleaegnifolium normal and nano for the management of downy mildew disease on cucumber caused by Pseudoperonospora cubensis under greenhouse conditions by reducing the incidence and severity of the disease and its reflection on the efficiency of the control and the area under the disease progress curve AUDPC compared to the fungicide Energy Previcur (Hcl - Propamocarb and Fosetyl-aluminum). The results of phenotypic and molecular diagnosis using polymerase chain reaction (PCR) technology for the two tested isolates showed that they are Pseudoperonospora cubensis. The nucleotide sequences were deposited in the gene bank under accession numbers ON509850 and ON509851. The results of the field trial indicated that the normal potassium silica at a concentration of 2.5 ml L-1 prevented the disease completely, with a severity rate and infection rate of 0.00%, it gave the highest reduction recorded with significant differences from the control treatment infected with artificial infestation 45.90% and 24.27%, respectively. They are followed by the normal and nano fungicide treatments. With regard to the effect of different treatments on the percentage of control, the results showed that the highest control percentage was recorded in the treatment of normal potassium silicates (100%), followed by the treatment of nano silicates (65.88%). Whereas, treatments of alcoholic and nano eggplant wild extract gave 51.64% and 50.50%, respectively, and 63.04 % and 63.09 % for the normal and nano fungicides, respectively.

Biocontrol of Sphaerotheca fuliginea, the causal agent of powdery mildew of cucumber by using aqueous extracts from five traditional Egyptian medicinal plants

Biocontrol of Sphaerotheca fuliginea, the causal agent of powdery mildew of cucumber by using aqueous extracts from five traditional Egyptian medicinal plants

Simulation of cucumber downy mildew Spread and sedimentation based on HYSPLIT

Cucumber downy mildew is a typical fungal airborne disease in which pathogenic spores are transmitted remotely by air currents, resulting in regional endemics. It is of great significance to study and analyze the temporal and spatial changes of downy mildew spores, especially the trajectory and sedimentation concentration of spores propagating with airflow in the air, for the prediction of downy mildew in cucumbers. Therefore, based on GDAS meteorological data from October to November 2017, this study uses the HYSPLIT-5 model to simulate the airflow propagation trajectory and sedimentation of downy mildew pathogen spores in the main cucumber planting areas in China, analyzes the trajectory frequency and sedimentation concentration in the long-distance and long-term series propagation process, and explores the transmission law of airborne disease spores, which provides a theoretical basis for predicting downy mildew in cucumbers.

Identification of Potential Phytochemical/Antimicrobial Agents against Pseudoperonospora cubensis Causing Downy Mildew in Cucumber through In-Silico Docking.

Compatibility interactions between the host and the fungal proteins are necessary to successfully establish a disease in plants by fungi or other diseases. Photochemical and antimicrobial substances are generally known to increase plant resilience, which is essential for eradicating fungus infections. Through homology modeling and in silico docking analysis, we assessed 50 phytochemicals from cucumber (Cucumis sativus), 15 antimicrobial compounds from botanical sources, and six compounds from chemical sources against two proteins of Pseudoperonospora cubensis linked to cucumber downy mildew. Alpha and beta sheets made up the 3D structures of the two protein models. According to Ramachandran plot analysis, the QNE 4 effector protein model was considered high quality because it had 86.8% of its residues in the preferred region. The results of the molecular docking analysis showed that the QNE4 and cytochrome oxidase subunit 1 proteins of P. cubensis showed good binding affinities with glucosyl flavones, terpenoids and flavonoids from phytochemicals, antimicrobial compounds from botanicals (garlic and clove), and chemically synthesized compounds, indicating the potential for antifungal activity.

Fungicidal spectrum and biological properties of a new fungicide, pyriofenone.

Pyriofenone is a new fungicide developed by Ishihara Sangyo Kaisha, Ltd. To determine the fungicidal spectrum of pyriofenone, in vivo pot tests and in vitro mycelial growth-inhibition tests were conducted. Pyriofenone showed excellent activity against wheat and cucumber powdery mildew and moderate efficacy against rice blast in the pot tests. In the mycelial growth-inhibition tests, most fungi were not affected by pyriofenone except for Botrytis cinerea, Helminthosporium sacchari, Pseudocercosporella herpotrichoides, Pyricularia oryzae, Rosellinia necatrix, and Verticillium dahliae. The fungicidal properties of pyriofenone on powdery mildew in cucumber and wheat were evaluated precisely. Pyriofenone exhibited excellent preventive and residual activities. It had high rainfastness in the cucumber leaves against powdery mildew. Pyriofenone also showed inhibitory activity on lesion development upon application until 2 days after inoculation, and the lesion expansion and sporulation of the cucumber powdery mildew fungus were effectively controlled. Furthermore, pyriofenone showed translaminar and vapor activities.

Triphenylphosphonium-Driven Targeting of Pyrimorph Fragment Derivatives Greatly Improved Its Action on Phytopathogen Mitochondria.

Pyrimorph is a carboxylic acid amide (CAA) fungicide, which shows excellent activity against oomycetes such as pepper phytophthora blight, tomato late blight, and downy mildew of cucumber. It works mainly by inhibiting the biosynthesis of cell wall of oomycetes. However, pyrimorph also shows weak activity of inhibiting mitochondrial complex III, which is the first CAA fungicide found to act on mitochondria. To improve this effect on mitochondria and develop fungicides that may have a novel mechanism of action, in this paper, by disassembling pyrimorph and conjugating the fragments with the mitochondrial-targeted delivery system (triphenylphosphonium), three series of mitochondrial-targeting analogues of pyrimorph were designed and synthesized. The results show that the pyridine-containing 1,1-diaryl is the core module of inhibition mitochondrial function of pyrimorph. Among these conjugates, compound 3b with a short linker showed the highest and broad-spectrum fungicidal activity, strong respiratory inhibition activity, and adenosine 5'-triphosphate synthesis inhibition activity, suggesting its potential as a fungicide candidate. 3b exhibited greatly improved action on mitochondria, such as by destroying the mitochondrial function of pathogens, causing mitochondrial swelling, weakening its influence on cell wall morphology, and so on. More importantly, this study provides a method to strengthen the drugs or pesticides with weak mitochondrial action, which is of special significance for developing mitochondrial bioactive molecules with the novel action mechanism.

Efficacy of iprovalicarb and copper oxychloride combination for the management of downy mildew of cucumber

Efficacy of iprovalicarb and copper oxychloride combination for the management of downy mildew of cucumber

Ultraviolet-B Irradiation Induces Resistance against Powdery Mildew in Cucumber (Cucumis sativus L.) through a Different Mechanism Than That of Heat Shock-Induced Resistance

Heat shock treatment (HST) and UV-B irradiation can reduce pathogen infection in crops. However, information on the mechanism of UV-B action is limited. Here, we investigated the mechanism of UV-B-induced resistance against powdery mildew in cucumber and compared it to that of heat-shock-induced resistance. We measured the percentage of leaf area showing disease symptoms and examined the expression levels of defense- and heat-shock-related genes across treatment groups. UV-B irradiation (intensity, 5 µW/cm2) for 4 h/d followed by pathogen inoculation reduced the appearance of powdery mildew by 21.17% compared with the control group. Unlike HST—which induces systemic resistance—UV-B irradiation induced local resistance in cucumber, as indicated by local changes in gene expression (Chi2 and ETR2). UV-B-treated plants inoculated with powdery mildew showed higher expression levels of Chi2, ETR2, and LOX6 than plants that were either treated with UV-B or inoculated. UV-B had no major effects on systemic acquired resistance or heat shock transcription factors, which are known to be affected by HST. Combined HST and UV-B had a strong synergistic effect in reducing powdery mildew in cucumber. Our results indicate that UV-B treatment likely operates through a different mechanism than HST in triggering cucumber resistance against powdery mildew infection.

Sigma factor binding protein 1 (CsSIB1) is a putative candidate of the major-effect QTL dm5.3 for downy mildew resistance in cucumber (Cucumis sativus).

The dm5.3 major-effect QTL in cucumber encodes a homolog of Arabidopsis sigma factor binding protein 1 (CsSIB1). CsSIB1 positively regulates defense responses against downy mildew in cucumber through the salicylic acid (SA) biosynthesis/signaling pathway. Downy mildew (DM) caused by the oomycete pathogen Pseudoperonospora cubensis is an important disease of cucumber and other cucurbits. Our knowledge on molecular mechanisms of DM resistance is still limited. In this study, we reported identification and functional characterization of the candidate gene for the major-effect QTL, dm5.3 for DM resistance originated from PI 197088. The dm5.3 QTL was Modelized through marker-assisted development of near isogenic lines (NILs). NIL-derived segregating populations were used for fine mapping which narrowed the dm5.3 locus down to a 144kb region. Based on multiple lines of evidence, we show that CsSIB1 (CsGy5G027140) that encodes the VQ motif-containing sigma factor binding protein 1 as the most likely candidate for dm5.3. Local association analysis identified a haplotype consisting of 7 SNPs inside the coding and promoter region of CsSIB1 that was associated with DM resistance. Expression of CsSIB1 was up-regulated with P. cubensis infection. Transcriptome profiling of NILs in response to P. cubensis inoculation revealed key players and associated gene networks in which increased expression of CsSIB1 antagonistically promoted salicylic acid (SA) but suppressed jasmonic acid (JA) biosynthesis/signaling pathways. Our work provides novel insights into the function of CsSIB1/dm5.3 as a disease resistance (R) gene. The roles of sigma factor binding protein genes in pathogen defense in cucumber were also discussed.

Residues, dissipation, and dietary risk assessment of oxadixyl and cymoxanil in cucumber

Oxadixyl and cymoxanil are widely used for controlling downy mildew in cucumber; however, there are few systematic studies on monitoring residue levels of these two pesticides in cucumber under greenhouse and open field conditions. In this study, a simplified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method was applied to analyze target compounds in cucumber. The average recoveries of oxadixyl and cymoxanil in cucumber ranged from 96% to 102%, with relative standard deviations (RSDs) of 1.8%–4.0%. The limits of quantification (LOQs) for two pesticides were both 0.01 mg/kg. The dissipation of oxadixyl was in accordance with a first-order kinetics equation, with half-lives ranging from 1.8 to 3.1 days. At the pre-harvest interval (PHI) of 3 or 5 days, the residue levels of oxadixyl in cucumber under open field conditions were higher than those under greenhouse conditions. Compared to oxadixyl, the cymoxanil degraded quickly, and its residues were below LOQ on the 3rd or 5th day after the last application. The terminal residues of oxadixyl and cymoxanil in the cucumber were both lower than the maximum residue limits (MRLs) in China. The risk quotient (RQ) used for dietary risk assessment was 1.8%–3.5% and 0.26%–0.51% for oxadixyl and cymoxanil, respectively. The results showed that the risks of these two pesticides used on cucumber at the experimental dosages are comparably acceptable for Chinese consumers of different gender and age groups. This study provides a reference data to use oxadixyl and cymoxanil scientifically and rationally.

Efficacy of Fungicides Used to Manage Downy Mildew in Cucumber Assessed with Multiple Meta-Analysis Techniques.

A nationwide, quantitative synthesis of fungicide efficacy data on management of cucurbit downy mildew (CDM) caused by Pseudoperonospora cubensis is needed to broadly evaluate fungicide performance. Three-level meta-analysis, three-level meta-regression, and network meta-analyses were conducted on data from 46 cucumber (Cucumis sativus) CDM fungicide efficacy studies conducted in the eastern United States retrieved from Plant Disease Management Reports published between 2009 and 2018. Three response variables were examined in each analysis: disease severity, marketable yield, and total yield, from which percent disease control and percent yield return compared with nontreated controls was calculated. Moderator variables used in the three-level meta-analysis or three-level meta-regression included year, disease pressure, number of fungicide applications, and slicing or pickling cucumbers. In the network meta-analysis, fungicides were grouped by common combinations of Fungicide Resistance Action Committee Codes and modes of action. Overall, fungicides significantly (P < 0.001) reduced disease severity and increased marketable and total yields, resulting in a mean 54.0% disease control and 61.9% marketable and 73.3% total yield return. Subgroup differences were observed for several fungicide applications, control plot disease severity, and cucumber type for marketable yield. Based on the meta-regression analysis for disease severity by year, fungicide efficacy has been decreasing from 2009 to 2018, potentially indicating broad development of fungicide resistance over time. Treatments containing quinone inside inhibitors, pyridinylmethyl-benzamides, and protectants and treatments containing oxysterol binding protein inhibitors and protectants most effectively reduced disease severity. The most effective fungicide combinations for disease control did not always result in the highest yield return.

Mechanisms of resistance to powdery mildew in cucumber

Podosphaera xanthii causes powdery mildew of cucumber, and is associated with significant yield and quality losses. Development of resistant or tolerant varieties is the most effective and eco-friendly strategy for powdery mildew management. An important host resistance mechanism is based on the recognition of conserved resistance genes, resulting in durable resistance. To determine powdery mildew resistance mechanisms in cucumber, total RNAs were isolated from the powdery mildew resistant cultivar Meltem, the tolerant line VT18, and the susceptible local variety Camlica. Expression levels of nine genes in these plants were analysed by Reverse Transcription Polymerase Chain Reaction (RT-PCR). The host reactions were assessed using microscope observations of stained specimens. Serine/threonine (STN7), transcription factor (WRKY22), serine/threonine-protein kinase (D6PKL1), and serine/threonine receptor kinase (NFP) genes were induced, as positive regulators in defence mechanisms against powdery mildew. Polygalacturonase Inhibitor (PGIP) did not express after P. xanthii inoculation of Camlica, resulting in susceptibility. After inoculation, callose synthase (CALLOSE) and cinnamyl alcohol dehydrogenase (CAD) gene expression levels were increased in resistant Meltem, but Hypersensitive Reaction (HR) and ROS formation were only linked in the tolerant VT18. Powdery mildew development was less in Meltem than in VT18, indicating that cell wall thickening and HR play separate roles in resistance to this disease.